Difference between revisions of "Team:Bielefeld-CeBiTec/Project/Mutation"

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            <div class="container text_header"><h1>Mutation</h1></div>
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<div class="container text_header"><h3> Overview </h3></div>
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            <div class="container text_header"><h2>Power through diversity</h2></div>
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      <br><br>
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      <div class="container text_header"><h3>Overview</h3></div>
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            One of the elementary requirements of every evolutionary process is variability, which can be achieved by
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            mutagenesis.
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            The alteration of existing genotypes is the only way to discover new features and abilities
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            of proteins and organisms. The aim of our mutagenesis system is to create a diversity of
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            binding proteins during the course of fermentation to gain as much different <a href="https://2016.igem.org/Team:Bielefeld-CeBiTec/Description">Evobodies </a>
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            as possible. Building on the library approach and leading to the selection system, the
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            mutagenesis represents the central part of our project. After transformation of the library and an initial selection round
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            the mutagenesis has to take over. That is why we are using a system that is capable of <i>in vivo</i>
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            DNA alteration. Two different approaches were applied to reach this aim.
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                    The mutagenesis plasmid is rather unspecific and leads to a general
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                raise of mutations in the DNA of host organisms. This system was used to get a manifold
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                increase of base pair exchanges and a strong repression of bacterial DNA repair mechanisms. Six
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                different proteins build this mutation system.
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                A more specific approach is the two plasmid mutagenesis system (Camps, 2003). It is
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                based on a modified DNA polymerase I, which replicates the first part of plasmids belonging
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                to the ColE1-family, like pSB1C3.
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            <br>
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            In the following, we will elaborate on why and how we implemented these two systems in detail in our project.
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Revision as of 11:00, 16 October 2016



Mutation

Power through diversity



Overview

One of the elementary requirements of every evolutionary process is variability, which can be achieved by mutagenesis. The alteration of existing genotypes is the only way to discover new features and abilities of proteins and organisms. The aim of our mutagenesis system is to create a diversity of binding proteins during the course of fermentation to gain as much different Evobodies as possible. Building on the library approach and leading to the selection system, the mutagenesis represents the central part of our project. After transformation of the library and an initial selection round the mutagenesis has to take over. That is why we are using a system that is capable of in vivo DNA alteration. Two different approaches were applied to reach this aim.
The mutagenesis plasmid is rather unspecific and leads to a general raise of mutations in the DNA of host organisms. This system was used to get a manifold increase of base pair exchanges and a strong repression of bacterial DNA repair mechanisms. Six different proteins build this mutation system.
A more specific approach is the two plasmid mutagenesis system (Camps, 2003). It is based on a modified DNA polymerase I, which replicates the first part of plasmids belonging to the ColE1-family, like pSB1C3.

In the following, we will elaborate on why and how we implemented these two systems in detail in our project.